Impacto do metilfenidato no estresse oxidativo e na plasticidade cerebral / Methylphenidate impact on oxidative stress and brain plasticity

Esse estudo avaliou o impacto do tratamento com cloridrato de metilfenidato (MFD) em crianças com Transtorno de Déficit de Atenção e Hiperatividade. O estudo incluiu metodologia variada, contendo estudo de revisão sobre efeito de metilfenidato sobre BDNF e estudo de coorte experimental. O estudo de revisão seguiu as diretrizes do PRISMA e foi registrado no PROSPERO. No estudo experimental, coorte aberta de centro único foi desenhada, com amostra de conveniência recrutada entre os anos de 2020 e 2022, no ambulatório de ensino da faculdade de Medicina da Universidade Federal de Viçosa (MG). Amostra de 62 crianças, 6 a 14 anos incompletos, sem tratamento prévio, diagnosticadas por psiquiatra infantil segundo os critérios do Manual Diagnóstico e Estatístico dos Transtornos Mentais (DSM5). Média de 8 consultas de acompanhamento clínico realizadas, com coletas de amostras biológicas em 3 delas: antes do início do MFD, com 12 e 24 semanas após uso da medicação. Amostra caracterizada quanto a dados sociodemográficos, sintomas de TDAH, avaliações clínica e psiquiátrica e testagem de inteligência pela psicologia. Amostras biológicas para dosagens séricas de marcadores oxidativos (níveis de capacidade antioxidante total -FRAP -, atividade de superóxido dismutase ­ SOD-, catalase ­ CAT -, glutathione -S-transferase -GST-, níveis de peroxidação lipídica e de proteínas carboniladas) foram coletadas de cada criança nos três momentos da avaliação. Metilfenidato de liberação imediata foi administrado na dosagem de média de 0,65mg/kg/dia. Usou-se o teste de Shapiro-Wilk e Kolmogorov-Smirnov para análise de normalidade. Frequências absolutas e relativas foram determinadas para as variáveis numéricas que foram descritas por suas médias e desvios padrões. Para comparações múltiplas dos parâmetros oxidativos foi realizado pós teste paramétrico de Tukey e para as demais variáveis análise de variância ANOVA (f). Análises dos parâmetros oxidativos foram realizadas no programa GraphPad Prism 7.0 (GraphPad Software, Inc. San Diego, CA, USA) e dos dados sociodemográficos e clínicos no software SPSS (versão 23.0 para Windows). Significância estatística foi considerada com p <0.05. Os resultados mostram: Sexo masculino predominante (71%), idade média 8,58 ± 1,91, predominância de apenas um cuidador - mãe e/ou pai biológico como chefe de família e maior frequência de tipo combinado de TDAH. Pressão arterial sistólica, frequência cardíaca e temperatura corporal alterações significativas, porém sem significância clínica. Índice massa corporal com diferença estatística, 37%, 19,3% e 21% das crianças apresentaram IMC acima do esperado para idade na avaliação 1, 2 e 3 respectivamente. Adesão ao tratamento medicamentoso permaneceu acima de 93,5% na 24ª semana. Durante o tratamento: FRAP não se alterou; atividade de SOD reduziu na 12ª semana em comparação à linha de base; atividade de CAT aumento significativo à 24ª em comparação 12ª semana; aumento significativo dos níveis de peroxidação lipídica à 24ª semana em comparação à 12ª semana. Aumento significativo das proteínas carboniladas na linha de base em comparação aos níveis da 12ª e 24ª semanas. O metilfenidato parece influenciar os parâmetros redox de crianças com TDAH, aumentando o estresse oxidativo. Porém, mecanismos cerebrais tamponam e desconhecemos o resultado dessas interações na estrutura cerebral. Níveis de BDNF não foram influenciados significativamente por metilfenidato em crianças com TDAH, quando comparados a controles em nossa metanálise.

This study evaluated the impact of methylphenidate hydrochloride (MFD) treatment (MFD) in children with Attention Deficit Hyperactivity Disorder. The study included varied methodology, including a review study about methylphenidate effects on BDNF and an experimental cohort study. The review study followed the PRISMA guidelines and was registered in PROSPERO. In the experimental study, a single-center open cohort was designed, with a convenience sample recruited between the years 2020 and 2022, at the teaching outpatient clinic of the Faculty of Medicine at Viçosa Federal University (UFV-MG). Sample with 62 children, 6 to 14 years old, without previous treatment, diagnosed by a child psychiatrist according to the criteria of the Diagnostic and Statistical Manual of Mental Disorders (DSM5). Eight clinical follow-up visits were carried out, and biological samples were collected in 3 visits: before MFD beginning and after 12- and 24-weeks medication. Sociodemographic data, ADHD symptoms, clinical and psychiatric assessments were performed, as well as intelligence testing by psychology. Biological samples for oxidative markers serum dosages (total antioxidant capacity levels -FRAP -, superoxide dismutase activity - SOD-, catalase - CAT -, glutathione S transferase -GST-, lipid peroxidation and carbonyl proteins levels) were collected of each child in the 3 evaluation moments. Immediate-release methylphenidate was administered at approximately 0.65mg/kg/day. The Shapiro-Wilk and Kolmogorov-Smirnov test was used for normality analysis. Absolute and relative frequencies were used for numeric variables that were described by their means and standard deviations. Tukey's parametric test and variance analysis ANOVA (f) were performed for multiple comparisons in redox parameters and other variables respectively. Redox parameters analysis was performed using GraphPad Prism 7.0 program (GraphPad Software, Inc. San Diego, CA, USA) and other variables using SPSS software (version 23.0 for Windows). Statistical significance was considered at p<0.05. Male was predominant (71%), with a mean age of 8.58 ± 1.91, mother and/or biological father were the householder in most homes. Systolic blood pressure, heart rate and body temperature had significant changes, but without clinical significance. Body mass index showed a statistical difference, with 37%, 19.3% and 21% of the children having a BMI above the expected for their age in assessment 1, 2 and 3 respectively. Combined-ADHD occurred in 58.1% of the children, inattentive in 32.3% and hyperactive/impulsive in 9.7%. Drug treatment adherence was 98.4% (12th week) and 93.5% (24th week). There were no changes in FRAP levels; SOD activity had significant decreased at week 12 compared to baseline activity; CAT activity showed a significant increase at the 24th week compared to 12th week; Significant increase in lipid peroxidation levels at 24th week compared to 12th week. Significant increase in protein carbonyls levels at baseline (before methylphenidate use) compared to levels at 12 and 24 weeks. Methylphenidate can influence the oxidative and antioxidative parameters of ADHD children, increasing oxidative stress. However, buffer brain mechanisms may act and the result of these interactions in brain structure is not completely known. BDNF levels were not significantly affected by methylphenidate treatment in ADHD children and do not differ from controls in our meta-analysis.

Virgin Coconut Oil Attenuates Deficits in Rats Undergoing Transient Cerebral Ischemia

@#Background and Objectives. Neuroprotection agents may help improve the outcomes of large vessel ischemic stroke. This study aims to explore the role of Virgin Coconut Oil (VCO), with its well-documented anti-oxidant properties, in neuroprotection after transient occlusion of the extracranial internal carotid artery in a rat model of stroke. Methods. Twenty-three Sprague-Dawley rats were randomized into two groups: 1) control group (n=11) given distilled water, and 2) treatment group (n=12) given virgin coconut oil at 5.15 ml/kg body weight for seven days. Subsequently, the rats underwent transient right extracranial internal carotid artery occlusion (EICAO) for 5 minutes using non-traumatic aneurysm clips. At 4 and 24 hours after EICAO, the animals were examined for neurologic deficits by an observer blinded to treatment groups, then sacrificed. Eight brain specimens (4 from each group) were subjected to histopathologic examination (H & E staining) while the rest of the specimens were processed using triphenyltetrazolium chloride (TTC) staining to determine infarct size and area of hemispheric edema. Results. VCO treatment significantly improved the severity of neurologic deficit (1.42 ± 2.31) compared to the control distilled water group (4.09 ± 2.59) 24 hours after EICAO. Whereas, infarct size and percent hemispheric edema did not significantly differ between the two groups. Conclusion. Prophylactic treatment of VCO is protective against EICAO-induced neurologic deficits in a rat model. VCO shows great potential as a neuroprotective agent for large vessel ischemic stroke. However, more studies are necessary to elucidate the neuroprotective mechanisms of VCO therapy in ischemic stroke.

Effect of pasteurization on the antioxidant and oxidant properties of human milk / Efeito da pasteurização nas propriedades antioxidantes e oxidantes do leite humano

ABSTRACT Objective: To evaluate the effect of pasteurization on antioxidant and oxidant properties of human milk. Methods: 42 samples of milk before and after pasteurisation were used to evaluate the antioxidant activity by the ferric reducing capacity and by scavenging the 2,2'-azino-bis 3-ethylbenzthiazoline-6-sulfonic acid radical. Lipid peroxidation was estimated by the concentration of malondialdehyde product using the thiobarbituric acid reactive substances assay and by the evaluation of advanced oxidation protein products. Results: No significant difference was observed in fresh human milk and after pasteurization in relation to antioxidant properties determined by the ferric reducing capacity (50.0±3.4% and 48.8±3.0%, respectively) and by scavenging the 2,2'-azino-bis 3-ethylbenzthiazoline-6-sulfonic acid radical (28.9±1.5% and 31.2±1.3%, respectively). The results of malondialdehyde (62.6±4.1 and 64.3±3.6 µM/mg) and protein oxidation products (59.4±3.4 and 54.2±3.8 µM/L) of fresh and pasteurized milk, respectively, did not exhibited any significant difference. Conclusions: This data showed that human milk has an important antioxidant activity and that the pasteurizing process does not influence the antioxidant capacity, avoiding the peroxidation of breast milk lipids and the formation of advanced protein oxidation products.

RESUMO Objetivo: Avaliar o efeito da pasteurização nas propriedades antioxidantes e oxidantes do leite humano. Métodos: Foram utilizadas 42 amostras de leite cru e após a pasteurização, para avaliação da atividade antioxidante pelos métodos da capacidade redutora do ferro e sequestro dos radicais derivados do ácido 2,2'-azino-bis (3-etilbenzotiazolina-6-sulfônico). A peroxidação lipídica (PL) foi estimada pela determinação das substâncias reativas ao ácido tiobarbitúrico e pela avaliação dos produtos proteicos de oxidação avançada. Resultados: Não se observou diferença significante no leite humano cru nem após a pasteurização em relação às propriedades antioxidantes determinadas pelo método da redução do ferro (50,0±3,4% e 48,8±3,0%, respectivamente) e pelo sequestro dos radicais derivados do ácido 2,2'-azino-bis (3-etilbenzotiazolina-6-sulfônico) (28,9±1,5% e 31,2±1,3%, respectivamente). Os resultados médios de malondialdeído [MDA] (62,6±4,1 e 64,3±3,6 µM/mg) e produtos de oxidação proteica (59,4±3,4 e 54,2±3,8 µM/L) entre os grupos leite fresco e leite pasteurizado, respectivamente, não evidenciaram diferença significante. Conclusões: Os dados demonstraram que o leite humano possui importante atividade antioxidante e que o processo de pasteurização não interfere nessa propriedade, evitando assim a peroxidação dos lipídios e a formação de produtos avançados de oxidação proteica.

Evaluation of serum prolidase enzyme activity and oxidative stress in patients with tinnitus / Avaliação da atividade da enzima sérica prolidase e do estresse oxidativo empacientes com zumbido

Abstract Introduction Tinnitus is defined as the perception of sound in the head or in the head in the absence of external sounds. The cause of tinnitus is still unknown. Objective We aimed to compare the serum levels of total oxidant status, total antioxidant status, serum prolidase enzyme activity and the oxidative stress index in patients with tinnitus to those of normal subjects. Methods Twenty five patients with tinnitus (mean age 34.3) and 25 healthy controls (mean age 37.2) were included in the study. Results Total oxidant status levels in the patient group were significantly higher than in the control group (p = 0.037). The mean total oxidant status value was 2.54 ± 0.95 mmoL/L in the patient group, and 2.06 ± 0.98 mmoL/L in the control group. The mean oxidative stress index level was 0.22 ± 0.10 AU in the patient group, while it was 0.17 ± 0.08 AU in the control group. Oxidative stress index was significantly higher in the patient group (0.026). There was no significant difference between the groups in terms of total antioxidant status values (p = 0.838). The mean serum prolidase enzyme activity level was 202.74 ± 33.56 U/L in the patient group and 175.46 ± 42.68 U/L in the control group. Serum prolidase enzyme activity levels in the patient group were significantly higher than in the control group (0.040). Conclusion We detected that the total oxidant status, oxidative stress index and serum prolidase enzyme activity levels were higher in patients with tinnitus when compared to the healthy controls. This finding suggests that oxidative stress index and serum prolidase enzyme activity may play a role in the etiopathogenesis of tinnitus.

Resumo Introdução O zumbido é definido como a percepção de um som na cabeça na ausência de sons externos. A causa do zumbido ainda é desconhecida. Objetivo Comparar os níveis séricos do estado oxidante total, estado antioxidante total, atividade da enzima prolidase sérica e o índice de estresse oxidativo em pacientes com zumbido e em indivíduos normais. Método Foram incluídos no estudo 25 pacientes com zumbido (média de 34,3 anos) e 25 controles saudáveis (média de 37,2 anos). Resultados Os níveis do estado oxidante total no grupo de pacientes foram significantemente maiores do que no grupo controle (p = 0,037). O valor médio do estado oxidante total foi de 2,54 ± 0,95 mmoL/L no grupo de pacientes e de 2,06 ± 0,98 mmoL/L no grupo controle. O nível médio de índice de estresse oxidativo foi de 0,22 ± 0,10 AU no grupo de pacientes, enquanto no grupo controle foi de 0,17 ± 0,08 AU. O índice de estresse oxidativo foi significantemente maior no grupo de pacientes (0,026). Não houve diferença significante entre os grupos em relação aos valores do estado antioxidante total (p = 0,838). O nível médio de atividade da enzima prolidase sérica foi de 202,74 ± 33,56 U/L no grupo de pacientes e de 175,46 ± 42,68 U/L no grupo controle. Os níveis de atividade da enzima prolidase sérica no grupo de pacientes foram significantemente maiores do que no grupo controle (0,040). Conclusão Detectamos que os níveis de estado oxidante total, índice de estresse oxidativo e atividade da enzima prolidase sérica foram maiores nos pacientes com zumbido quando comparados aos controles saudáveis. Esse achado sugere que o índice de estresse oxidativo e a atividade da enzima prolidase sérica podem desempenhar um papel na etiopatogenia do zumbido.

Possible protective effect of olive leaves extract on paracetamol induced hepatotoxicity in male albino rats / Possível efeito protetor do extrato de folhas de oliveira na hepatotoxicidade induzida por paracetamol em ratos albinos machos

Paracetamol (PCM) overdose can cause hepatotoxicity with oxidative stress; the present study was carried out to establish the possible protective effect of olive leaves extract (OLE) on toxicity induced by paracetamol in adult male rats. Twenty four adult male rats were divided into four equal groups; control, olive leaves extract group, paracetamol group and olive leaves extract plus paracetamol group. Some biochemical parameters and liver histopathology were evaluated. PCM treatment significantly increased serum aspartate aminotransferase (AST), alanine aminotransferase (ALT), total bilirubin, gamma-glutamyltransferase (GGT), lactate dehydrogenase (LDH), urea, creatinine and alpha-fetoprotein. Paracetamol was found to significantly increase malonaldehyde (MDA) and decrease glutathione reductase (GR) activity in tissue and significantly decrease total antioxidant capacity (TAC) and superoxide dismutase (SOD) in serum. Administration of OLE caused a significant decrease serum AST, ALT enzyme, total bilirubin, GGT, LDH, creatinine, urea, alpha-fetoprotein. Also, amelioration of oxidant ­ antioxidant status with olive leaves extract was observed in addition to a significant decrease in MDA and a significant increase in TAC in liver tissue with a significant increase in glutathione reductase (GR) and SOD in serum compared to paracetamol treated group The chemical pathological changes were in step with histopathological observation suggesting marked hepatoprotective result of olive leaves extract. It could be concluded that olive leaves extract (OLE) treatment may be effective in decreasing hepatic injury and oxidative stress induced by paracetamol overdose in male albino rats

A sobredosagem de paracetamol (PCM) pode causar hepatotoxicidade com estresse oxidativo; o presente estudo foi realizado para estabelecer o possível efeito protetor do extrato de folhas de oliveira (OLE) na toxicidade induzida pelo paracetamol em ratos machos adultos. Vinte e quatro ratos machos adultos foram divididos em quatro grupos iguais: controle, grupo extrato de folhas de oliveira, grupo paracetamol e extrato de folhas de oliveira mais grupo paracetamol. Alguns parâmetros bioquímicos e histopatologia hepática foram avaliados. O tratamento com PCM aumentou significativamente aspartato aminotransferase sérica (AST), alanina aminotransferase (ALT), bilirrubina total, gama-glutamiltransferase (GGT), lactato desidrogenase (LDH), uréia, creatinina e alfa-fetoproteína. Verificou-se que o paracetamol aumenta significativamente o malonaldeído (MDA) e diminui a atividade da glutationa redutase (GR) no tecido e diminui significativamente a capacidade antioxidante total (TAC) e a superóxido dismutase (SOD) no soro. A administração de OLE causou uma diminuição significativa de AST, enzima ALT, bilirrubina total, GGT, LDH, creatinina, uréia, alfa-fetoproteína. Também foi observada melhora do status oxidante - antioxidante com extrato de folhas de oliveira, além de uma diminuição significativa no MDA e um aumento significativo no TAC no tecido hepático, com um aumento significativo na glutationa redutase (GR) e SOD no soro em comparação ao grupo tratado com paracetamol. As alterações patológicas químicas acompanharam a observação histopatológica, sugerindo resultado hepatoprotetor acentuado do extrato de folhas de oliveira. Pode-se concluir que o tratamento com extrato de folhas de oliveira (OLE) pode ser eficaz na diminuição da lesão hepática e do estresse oxidativo induzido pela overdose de paracetamol em ratos albinos machos

Correlación entre los microorganismos oxidantes del amonio y los factores ambientales durante el compostaje de estiércol de ganado / Correlation between ammonia-oxidizing microorganisms and environmental factors during cattle manure composting

Abstract Cattle manure composting was performed in an aerated vessel. Community structure and diversity of ammonia-oxidizing bacteria (AOB) and ammonia-oxidizing archaea (AOA) were investigated using polymerase chain reaction and denaturing gradient gel electrophoresis (PCR-DGGE) techniques targeting the ammonia monooxygenase alpha subunit (amoA) gene and the correlation between AOB and AOA communities and environmental factors was explored. Thirteen (13) AOB sequences were obtained, which were closely related to Nitrosomonas spp., Nitrosomonas eutropha, and Nitrosospira spp. and uncultured bacteria, among which Nitrosomonas spp. were predominant. Excessively high temperature and high ammonium concentration were not favorable for AOB growth. Five AOA sequences, belonging to Candidatus Nitrososphaera gargensis and to an uncultured archaeon, were obtained. During composting, community diversity of AOB and AOA fluctuated, with AOA showing a higher Shannon-Wiener index. The AOB community changed more dramatically in the mesophilic stage and the early thermophilic stage, whereas the most obvious AOA community succession occurred in the late thermophilic stage, the cooling stage and the maturity stage. Water content, total nitrogen (TN) and ammonium concentration were more relevant to the AOB community structure, while higher correlations were observed between ammonia, nitrate and TN and the AOA community. AOB community diversity was negatively correlated with pH (r = -0.938, p < 0.01) and water content (r = -0.765, p < 0.05), while positively correlated with TN (r = 0.894, p < 0.01). AOA community diversity was negatively correlated with ammonium concentration (r = -0.901, p < 0.01). Ammonium concentration played an important role in the succession of AOB and AOA communities during composting.

Resumen Se llevó a cabo un compostaje de estiércol de ganado en un recipiente aireado. Se investigó la estructura de la comunidad y la diversidad de bacterias oxidantes del amoníaco (AOB) y las arqueas oxidantes del amoníaco (AOA) mediante el uso de las técnicas de reacción en cadena de la polimerasa y la electroforesis en gel con gradiente de desnaturalización (PCR-DGGE) dirigidas al gen de la subunidad alfa de la amonio monooxigenasa (amoA), y se exploró la correlación entre las comunidades AOB, AOA y los factores ambientales. Se obtuvieron 13 secuencias de AOB, las cuales se relacionaron estrechamente con Nitrosomonas spp., Nitrosomonas eutropha y Nitrosospira spp., y bacterias no cultivadas, entre las cuales fueron predominantes las Nitrosomonas spp. La temperatura excesivamente alta y la concentración de amonio elevada no fueron favorables para el crecimiento de las AOB. Se obtuvieron 5 secuencias de AOA, pertenecientes a Candidatus Nitrososphaera gargensis y un Archaeon no cultivado. Durante el compostaje, la diversidad de AOB y AOA fluctuó y las AOA mostraron un índice de Shannon-Wiener más alto. La comunidad de AOB cambió significativamente en la etapa mesofílica y la etapa termofílica temprana, mientras que la sucesión más obvia de la comunidad AOA ocurrió en la etapa termofílica tardía y las etapas de enfriamiento y de maduración. El contenido de agua, el nitrógeno total (TN) y la concentración de amonio fueron más relevantes para la estructura de la comunidad AOB, mientras que se observaron correlaciones mayores entre amoníaco, nitrato y TN, y la comunidad AOA. La diversidad de la comunidad AOB se correlacionó negativamente con el pH (r= -0,938; p < 0,01) y el contenido de agua (r = -0,765; p < 0,05), mientras que se relacionó positivamente con TN (r = 0,894; p < 0,01). La diversidad de la comunidad AOA se correlacionó negativamente con la concentración de amonio (r = -0,901; p < 0,01). La concentración de amonio desempenó un papel importante en la sucesión de las comunidades AOB y AOA durante el compostaje.

Investigating the effect of intracameral cefuroxime on oxidative stress and apoptosis in the rat cornea / Investigando o efeito de cefuroxima intracameral sobre o estresse oxidativo e a apoptose na córnea de ratos

ABSTRACT PURPOSE: We examined the effect of intracameral administration of cefuroxime on oxidative stress and endothelial apoptosis in rat corneal tissue. METHODS: In total, 30 rats were divided into 3 groups of 10 rats each (intracameral administration of cefuroxime 0.1 mg/0.01 mL (cefuroxime group); intracameral administration of balanced salt solution 0.01 mL (control group); or absence of intracameral injection (sham group). Corneal endothelial apoptosis was assessed by immunohistochemical analysis using caspase-3 and caspase-8. Total oxidant status, total antioxidant status, oxidative stress index, and paraoxonase and arylesterase levels were examined in corneal endothelial tissue and serum. RESULTS: Paraoxonase levels in the serum were significantly different between the sham and cefuroxime groups (p=0.027). A significant difference was also observed in total oxidant status levels between the cefuroxime and balanced salt solution groups (p=0.023). In addition, there were significant differences in total antioxidant status levels in corneal tissue between the cefuroxime and sham groups (p<0.001) and between the cefuroxime and balanced salt solution groups (p<0.001). Furthermore, significant differences were also observed in oxidative stress index levels between the cefuroxime and balanced salt solution groups (p=0.001) and between the cefuroxime and sham groups (p=0.026). According to the immunohistochemical staining results, a significant association with caspase-3 activity existed between the cefuroxime and balanced salt solution groups (p=0.007), while no significant difference was found with caspase-8 activity (p=0.541). Caspase-3 activity exhibited a significant relationship between the sham and balanced salt solution groups (p=0.018), but no relationship was found with caspase-8 activity (p=0.623). CONCLUSION: Immunohistochemical examination revealed that intracameral cefuroxime increased apoptosis when compared to the sham and balanced salt solution groups. Moreover, intracameral cefuroxime increased oxidative stress in the cornea and simultaneously induced apoptosis.

RESUMO OBJETIVO: Examinamos o efeito da administração intracameral da cefuroxima sobre o estresse oxidativo e a apoptose endotelial no tecido corneano de ratos. MÉTODOS: No total, 30 ratos foram divididos em 3 grupos de 10 ratos cada (administração intracameral de cefuroxima 0,1 mg/0,01 mL (grupo cefuroxima), administração intracameral de solução salina balanceada 0,01 mL (grupo controle) ou ausência de injeção intracameral (grupo sham)). A apoptose endotelial da córnea foi avaliada por análise imuno-histoquimica usando caspase-3 e -8. O status oxidante total, o status antioxidante total, o índice de estresse oxidativo e os níveis de a paraoxonase e arilesterase foram investigados no tecido endotelial da córnea e no soro. RESULTADOS: Os níveis de paraoxonase no soro foram significativamente diferentes entre os grupos sham e cefuroxima (p=0,027). Foi também observada uma diferença significativa nos níveis de estado oxidante total entre os grupos cefuroxima e solução salina balanceada (p=0,023). Além disso, houve diferenças significativas nos níveis de status antioxidante total no tecido da córnea entre os grupos cefuroxima e sham (p<0,001) e entre os grupos cefuroxima e solução salina balanceada (p<0,001). Diferenças significativas também foram observadas nos níveis do índice de estresse oxidativo entre os grupos cefuroxima e solução salina balanceada (p=0,001) e entre os grupos cefuroxima e sham (p=0,026). De acordo com os resultados de coloração imuno-histoquimica, houve associação significativa com a atividade da caspase-3 entre os grupos cefuroxima e solução salina balanceada (p=0,007), enquanto não houve diferença significativa com a atividade da caspase-8 (p=0,541). A atividade da caspase-3 exibiu uma relação significativa entre os grupos sham e solução salina balanceada (p=0,018), mas nenhuma relação foi encontrada com a atividade da caspase-8 (p=0,623). CONCLUSÃO: O exame imuno-histoquímico revelou que a cefuroxima intracameral aumentou a apoptose quando comparada com os grupos sham e solução salina balanceada. Além disso, a cefuroxima intracameral aumentou o estresse oxidativo na córnea e induziu simultaneamente a apoptose.

Monitoring Glutathione Dynamics and Heterogeneity in Living Stem Cells

Glutathione (GSH) is a major antioxidant in cells, and plays vital roles in the cellular defense against oxidants and in the regulation of redox signals. In a previous report, we demonstrated that stem cell function is critically affected by heterogeneity and dynamic changes in cellular GSH concentration. Here, we present a detailed protocol for the monitoring of GSH concentration in living stem cells using FreSHtracer, a real-time GSH probe. We describe the steps involved in monitoring GSH concentration in single living stem cells using confocal microscopy and flow cytometry. These methods are simple, rapid, and quantitative, and able to demonstrate intracellular GSH concentration changes in real time. We also describe the application of FreSHtracer to the sorting of stem cells according to their GSH content using flow cytometry. Typically, microscopic or flow cytometric analyses of FreSHtracer and MitoFreSHtracer signals in living stem cells take ~2~3 h, and the fractionation of stem cells into subpopulations on the basis of cellular GSH levels takes 3~4.5 h. This method could be applied to almost every kind of mammalian cell with minor modifications to the protocol described here.

Male Oxidative Stress Infertility (MOSI): Proposed Terminology and Clinical Practice Guidelines for Management of Idiopathic Male Infertility

Despite advances in the field of male reproductive health, idiopathic male infertility, in which a man has altered semen characteristics without an identifiable cause and there is no female factor infertility, remains a challenging condition to diagnose and manage. Increasing evidence suggests that oxidative stress (OS) plays an independent role in the etiology of male infertility, with 30% to 80% of infertile men having elevated seminal reactive oxygen species levels. OS can negatively affect fertility via a number of pathways, including interference with capacitation and possible damage to sperm membrane and DNA, which may impair the sperm's potential to fertilize an egg and develop into a healthy embryo. Adequate evaluation of male reproductive potential should therefore include an assessment of sperm OS. We propose the term Male Oxidative Stress Infertility, or MOSI, as a novel descriptor for infertile men with abnormal semen characteristics and OS, including many patients who were previously classified as having idiopathic male infertility. Oxidation-reduction potential (ORP) can be a useful clinical biomarker for the classification of MOSI, as it takes into account the levels of both oxidants and reductants (antioxidants). Current treatment protocols for OS, including the use of antioxidants, are not evidence-based and have the potential for complications and increased healthcare-related expenditures. Utilizing an easy, reproducible, and cost-effective test to measure ORP may provide a more targeted, reliable approach for administering antioxidant therapy while minimizing the risk of antioxidant overdose. With the increasing awareness and understanding of MOSI as a distinct male infertility diagnosis, future research endeavors can facilitate the development of evidence-based treatments that target its underlying cause.

Evaluating clinical and laboratory effects of ozone in non-surgical periodontal treatment: a randomized controlled trial

Abstract Objective: This study aims to evaluate the clinical and biochemical (oxidative stress and pro-inflammatory mediators) effects of the gaseous ozone use accompanied by scaling and root planning (SRP) in periodontal treatment. Material and Methods: The study population consisted of 40 patients with chronic periodontitis (CP) randomly sorted into two groups of 20. The experimental group received SRP plus 3 watts gaseous ozone in two separate applications five days apart, whereas the control group received SRP plus placebo. Clinical periodontal parameters were assayed and saliva samples were taken before the initial and one month after the second treatment. Periodontal examination assessed plaque index (PI), gingival index (GI), probing depth, and clinical attachment level (CAL). Total antioxidant status (TAS), total oxidant status (TOS), nitric oxide (NO), 8-hydroxy-2'-deoxyguanosine (8-OHdG), myeloperoxidase (MPO), glutathione (GSH), malondialdehyde (MDA), and transforming growth factor-beta (TGF-β) levels were evaluated from saliva samples. Results: Changes following treatment in PI, GI, probing depth, and CAL scores were similar for both groups (p>0.05). Of note, TGF-β levels were observed to be higher in the treatment group than in controls (p<0.05). Changes in 8-OHdG, TAS, TOS, NO, MPO, GSH and MDA levels, however, were not significantly different between groups (p>0.05). Conclusion: The findings of this study indicate that SRP plus gaseous ozone versus SRP alone does not correlate to a significant improvement in periodontal recovery.

Antithrombotic effect of epigallocatechin gallate on the patency of arterial microvascular anastomoses

BACKGROUND: Microvascular anastomosis patency is adversely affected by local and systemic factors. Impaired intimal recovery and endothelial mechanisms promoting thrombus formation at the anastomotic site are common etiological factors of reduced anastomosis patency. Epigallocatechin gallate (EGCG) is a catechin derivative belonging to the flavonoid subgroup and is present in green tea (Camellia sinensis). This study investigated the effects of EGCG on the structure of vessel tips used in microvascular anastomoses and evaluated its effects on thrombus formation at an anastomotic site. METHODS: Thirty-six adult male Wistar albino rats were used in the study. The right femoral artery was cut and reanastomosed. The rats were divided into two groups (18 per group) and were systemically administered either EGCG or saline. Each group were then subdivided into three groups, each with six rats. Axial histological sections were taken from segments 1 cm proximal and 1 cm distal to the microvascular anastomosis site on days 5, 10, and 14. RESULTS: Thrombus formation was significantly different between the EGCG and control groups on day 5 (P=0.015) but not on days 10 or 14. The mean luminal diameter was significantly greater in the EGCG group on days 5 (P=0.002), 10 (P=0.026), and 14 (P=0.002). Intimal thickening was significantly higher on days 5 (P=0.041) and 10 (P=0.02). CONCLUSIONS: EGCG showed vasodilatory effects and led to reduced early thrombus formation after microvascular repair. Similar studies on venous anastomoses and random or axial pedunculated skin flaps would also contribute valuable findings relevant to this topic.

Tratamento clínico de zumbido primário em adultos e idosos: revisão sistemática / Clinical treatment of primary tinnitus in adults and in the elderly: systematic review

O zumbido no ouvido é definido como uma ilusão auditiva ou sensação sonora endógena, não relacionada a nenhuma fonte externa de estimulação. É um sintoma frequente na população idosa. Até hoje, vários autores argumentam que o desconhecimento da etiologia do zumbido, aliado à subjetividade desta manifestação, mais a sobreposição das enfermidades e dos sintomas que, geralmente, acometem os pacientes idosos, dificultam a obtenção de um bom resultado terapêutico. O objetivo desta revisão foi levantar quais os tratamentos clínicos mais utilizados na prática clínica no tratamento do zumbido primário em adultos e idosos. Procedeu-se à verificação do status dos últimos 5 anos de estudos em textos de acesso livre, no banco de dados eletrônicos da PubMed. Apresentaram tratamentos clínicos para o zumbido primário 25 artigos; aqueles com resultados satisfatórios foram quatro artigos sobre acupuntura, dois sobre neuromodulação de resenha coordenada acústica, um sobre uso combinado de amplificação e gerador de som, e um sobre psicoterapia corporal, que incluíam tanto adultos e idosos, tendo a idade média entre 51 a 54 anos. Não se pode afirmar que os tratamentos propostos são eficazes na cura dos sintomas de zumbido em adultos e idosos, mas sim que existem algumas terapêuticas de baixo custo que apresentam respostas relativamente satisfatórias. (AU)

Tinnitus is defined as a hearing illusion or endogenous auditory sensation that is not related to any external stimulation source. It is a frequent symptom among elderly people. To date, many authors have argued that the lack of knowledge about the tinnitus etiology, added to the subjectivity of this manifestation, and the overlap of other diseases and symptoms that often occur with aged patients make the obtainment of a good therapeutic result difficult. The objective of this review was to find the most used clinical treatment in clinical practice for primary tinnitus on adults and elderly. The status of the last five years of studies in free full texts on PubMed database was checked. Twenty-five articles showed clinical treatment for primary tinnitus, with four articles about acupuncture, two about acoustic coordinate reset neuromodulation, one about sound generator associated with conventional amplification, and one about body-psychotherapy which included adults and elderly with an average age between 51 to 54 years old showing satisfactory results. It is difficult to state that the proposed treatment is efficient on healing the tinnitus symptoms on adults and elderly but there are some low-cost therapies showing relatively satisfactory responses. (AU)

Effect of wheatgrass on DNA damage, oxidative stress index and histological findings in diabetic rats / Efecto del pasto de trigo sobre el daño del ADN, el índice de estrés oxidativo y los hallazgos histológicos en ratas diabéticas

This study was aimed to search the effect of wheatgrass on the Total Antioxidan (TAS)-Oxidan Status (TOS) and DNA damage in rat with diabetes. The rats used in the study were randomly divided into 4 groups that each of has 10 rats: Control group; 1 ml single dose phosphate-citrate buffer injected i.p (pH: 4.5), Diabetes group; 45 mg/kg single dose streptozotocin injected i.p., Wheatgrass group; was given oral wheatgrass (10 ml/kg/day) for 6 weeks, Diabetes +Wheatgrass group; 45 mg/kg single dose streptozotocin injected i.p. and wheatgrass (10 ml/kg/day) was given by oral during 6 weeks. After the process of experiment during 6 weeks, blood sample and pancreas tissue were taken. The analysis were done of blood glucose levels, TAS, TOS levels by colorimetric kits; DNA damage by ELISA kits in serum. The pancreas tissues were examined histopathologically. In the group of Diabetes+Wheatgrass was determined that the levels of glucose levels (p<0.001), TOS (p<0.05) and OSI (p<0.01) statistically decreased and heal histopatolojical compared to diabetes group. In the group of Wheatgrass was determined that the levels of TAS p<0.05 statistically increased from other groups. The statistical significance were not found in the level of serum 8OHdG differences between the groups. The beta cells were seen to increase in the group receiving wheatgrass for therapeutic purposes.As a conclusion, it was determined that wheatgrass strengthened the anti-oxidant defense system and reduced the glucose level in diabetic rats.

El objetivo de este estudio fue buscar el efecto del pasto de trigo sobre el estado total de antioxidantes (TAS) -Oxidan Status (TOS) y el daño del ADN en ratas con diabetes. Las ratas analizadas en el estudio se dividieron aleatoriamente en 4 grupos de 10 ejemplares cada uno: grupo control; 1 ml de tampón fosfato-citrato de dosis única inyectado i.p. (pH: 4,5)., Grupo diabetes; 45 mg / kg de estreptozotocina en dosis única inyectada i.p., grupo pasto de trigo; se administró pasto de trigo oral (10 ml / kg / día) durante 6 semanas, grupo diabetes + pasto de trigo; 45 mg / kg de estreptozotocina en dosis única inyectada i.p. y pasto de trigo (10 ml / kg / día) por vía oral durante 6 semanas. Después del proceso experimental durante 6 semanas, se tomaron muestras de sangre y tejido de páncreas. Se midieron los niveles de glucosa en sangre, TAS, y TOS mediante kits colorimétricos; El daño al ADN fue realizado por kits de ELISA en suero. Los tejidos del páncreas se examinaron histopatológicamente. En el grupo de diabetes + pasto de trigo se determinó que los niveles de glucosa (p <0,001), TOS (p <0,05) y OSI (p <0,01) disminuyeron estadísticamente y curaron histopatológicamente en comparación con el grupo de diabetes. En el grupo de pasto de trigo se determinó que los niveles de TAS p <0,05 se incrementaron estadísticamente con respecto a otros grupos. No fue estadísticamente significativo el nivel de las diferencias séricas de 8OHdG entre los grupos. Se observó que las células beta aumentaron en el grupo que recibió pasto de trigo con fines terapéuticos. Como conclusión, se determinó que el pasto de trigo fortaleció el sistema de defensa antioxidante y redujo el nivel de glucosa en las ratas diabéticas.

The Diagnostic Value of Malondialdehyde, Superoxide Dismutase and Catalase Activity in Drug Naïve, First Episode, Non-Smoker Generalized Anxiety Disorder Patients

OBJECTIVE: Generalized anxiety disorder (GAD) is a common anxiety disorder. Although lots of research done to reveal neurobiological basis of GAD, it is still unclear. Diagnosis of GAD depends on subjective complaints of patients, thus the need for a biological marker is constantly emerging. In this study, we aimed to investigate diagnostic value of malondialdehyde (MDA), superoxide dismutase (SOD) and catalase (CAT) in GAD. METHODS: We evaluated MDA, SOD, and CAT levels in peripheral blood of 46 patients and 45 controls. MDA was measured with Ohkawa’s methods, SOD was measured with Fridovich method, and CAT was measured with Beutler’s method. RESULTS: MDA was significantly increased in patients than controls, medians 4.05 nmol/mg and 1.71 nmol/mg respectively, p < 0.001; SOD and CAT activity was significantly decreased in patients than controls, medians of SOD were 159.07 U/mg and 301.87 U/mg, p < 0.001 respectively, medians for CAT were 138.47 U/mg and 160.60 U/mg respectively. We found high correlation between Hamilton Anxiety Rating Scale and SOD, MDA r values were 0.723 and 0.715 respectively, p < 0.001 for both. Receiver operator characteristic (ROC) curve analysis showed high diagnostic performance for MDA and SOD, low diagnostic performance for CAT, areas under curve were 1.0, 1.0, and 0.648 respectively. CONCLUSION: Our results reveal possible diagnostic value of MDA, less likely of SOD but not CAT. Future studies should investigate diagnostic value of oxidants and antioxidantn enzymes in larger samples and include diagnostic value of these parameters.

Review of Disinfection and Sterilization – Back to the Basics

In-depth knowledge of disinfection and sterilization is a key component of infection control. Sterilization completely removes a spore, whereas disinfection cannot. Disinfectants are classified as oxidants and non-oxidants. The decision regarding which method to apply is based on Spaulding's classification. In this article, disinfection and sterilization are thoroughly reviewed, and extensive information from basic to practical points is discussed.

Review of Disinfection and Sterilization – Back to the Basics

In-depth knowledge of disinfection and sterilization is a key component of infection control. Sterilization completely removes a spore, whereas disinfection cannot. Disinfectants are classified as oxidants and non-oxidants. The decision regarding which method to apply is based on Spaulding's classification. In this article, disinfection and sterilization are thoroughly reviewed, and extensive information from basic to practical points is discussed.

Ursolic acid in health and disease

Ursolic acid (UA) is a natural triterpene compound found in various fruits and vegetables. There is a growing interest in UA because of its beneficial effects, which include anti-inflammatory, anti-oxidant, anti-apoptotic, and anti-carcinogenic effects. It exerts these effects in various tissues and organs: by suppressing nuclear factor-kappa B signaling in cancer cells, improving insulin signaling in adipose tissues, reducing the expression of markers of cardiac damage in the heart, decreasing inflammation and increasing the level of anti-oxidants in the brain, reducing apoptotic signaling and the level of oxidants in the liver, and reducing atrophy and increasing the expression levels of adenosine monophosphate-activated protein kinase and irisin in skeletal muscles. Moreover, UA can be used as an alternative medicine for the treatment and prevention of cancer, obesity/diabetes, cardiovascular disease, brain disease, liver disease, and muscle wasting (sarcopenia). In this review, we have summarized recent data on the beneficial effects and possible uses of UA in health and disease managements.

The Relationship between the Number of Manic Episodes and Oxidative Stress Indicators in Bipolar Disorder

OBJECTIVE: Bipolar disorder (BD) is a chronic mood disorder characterized by recurrent episodes that has a lifetime prevalence of 0.4–5.5%. The neurochemical mechanism of BD is not fully understood. Oxidative stress in neurons causes lipid peroxidation in proteins associated with neuronal membranes and intracellular enzymes and it may lead to dysfunction in neurotransmitter reuptake and enzyme activities. These pathological processes are thought to occur in brain regions associated with affective functions and emotions in BD. The relationship between the number of manic episodes and total oxidant-antioxidant capacity was investigated in this study. METHODS: Eighty-two BD patients hospitalized due to manic symptoms and with no episodes of depression were enrolled in the study. Thirty of the 82 patients had had their first episode of mania, and the other 52 patients had had two or more manic episodes. The control group included 45 socio-demographically matched healthy individuals. Serum total antioxidant capacity (TAC) and total oxidant capacity (TOC) measurements of the participants were performed. The oxidative stress index (OSI) was calculated by TOC/TAC. RESULTS: There were no significant differences in OSI scores between BD patients with first-episode mania and BD patients with more than one manic episode. However, OSI scores in both groups were significantly higher than in the control group. TOC levels of BD patients with first-episode mania were found to be significantly higher than TOC levels of BD patients with more than one manic episode and healthy controls. There were no significant differences in TAC levels between BD patients with first-episode mania and BD patients with more than one manic episode. TAC levels in both groups were significantly higher than in the control group. CONCLUSION: Significant changes in oxidative stress indicators were observed in this study, confirming previous studies. Increased levels of oxidants were shown with increased disease severity rather than with the number of manic episodes. Systematic studies, including of each period of the disorder, are needed for using the findings indicating deterioration of oxidative parameters.

Differential expression on mitochondrial tryparedoxin peroxidase (mTcTXNPx) in Trypanosoma cruzi after ferrocenyl diamine hydrochlorides treatments

Abstract Resistance to benznidazole in certain strains of Trypanosoma cruzi may be caused by the increased production of enzymes that act on the oxidative metabolism, such as mitochondrial tryparedoxin peroxidase which catalyses the reduction of peroxides. This work presents cytotoxicity assays performed with ferrocenyl diamine hydrochlorides in six different strains of T. cruzi epimastigote forms (Y, Bolivia, SI1, SI8, QMII, and SIGR3). The last four strains have been recently isolated from triatominae and mammalian host (domestic cat). The expression of mitochondrial tryparedoxin peroxidase was analyzed by the Western blotting technique using polyclonal antibody anti mitochondrial tryparedoxin peroxidase obtained from a rabbit immunized with the mitochondrial tryparedoxin peroxidase recombinant protein. All the tested ferrocenyl diamine hydrochlorides were more cytotoxic than benznidazole. The expression of the 25.5 kDa polypeptide of mitochondrial tryparedoxin peroxidase did not increase in strains that were more resistant to the ferrocenyl compounds (SI8 and SIGR3). In addition, a 58 kDa polypeptide was also recognized in all strains. Ferrocenyl diamine hydrochlorides showed trypanocidal activity and the expression of 25.5 kDa mitochondrial tryparedoxin peroxidase is not necessarily increased in some T. cruzi strains. Most likely, other mechanisms, in addition to the over expression of this antioxidative enzyme, should be involved in the escape of parasites from cytotoxic oxidant agents.

Increased antioxidant activity after exposure of ozone in murine asthma model

BACKGROUND: Ozone is well known as an important component of ambient air pollutants. Ozone can aggravate respiratory symptoms in patients with bronchial asthma, but, not in healthy person. We hypothesized asthma itself may show different response to ozone compared to nonasthma. OBJECTIVE: This study was performed to evaluate the differences of response to ozone between normal and asthmatic mice model in terms of status of oxidant injury and antioxidant activity. METHODS: Three parts per million of ozone was exposed to ovalbumin (OVA)-induced murine asthma model for 3 hours at 3, 7, 14, 21 days after completion of asthma model. Airway responsiveness to methacholine was measured after completion of asthma model. Bronchoalveolar lavage (BAL), protein extraction from lung for Western blot and immunohistochemistry of 4-hydroxy-2-nonenal (4-HNE), proliferating cell nuclear antigen (PCNA), NF-E2 related factor 2 (Nrf-2), and activity of glutathione were performed at before and each ozone exposure day. RESULTS: Airway hyper-responsiveness and increased eosinophils in BAL fluid were observed in asthma model. In asthma model, the expression of 4-HNE already more increased at baseline (without ozone) compared to those in sham model. This increased expression is more enhanced at 3 days after ozone exposure. The expression of PCNA was significantly increased in OVA-model compared to those in sham model. The expression of Nrf-2 was observed at baseline, and 3 and 7 days after exposure ozone in asthma model, but not in sham model. The activity of glutathione increased significantly after exposure of ozone, but not in sham model. CONCLUSION: Murine asthma model has enhanced oxygen toxicity and antioxidant activity response to ozone.